Mouse kidney cortex or podocytes were homogenized in lysis buffer. After concentration being measured, protein was subjected to SDS-PAGE gel under reducing condition and transferred onto a nitrocellulose membrane. After blocking with 5% milk, the membrane was incubated with anti-GAPDH (Millipore), anti-TSP1 (Thermo Scientific), anti-WT1 (Santa Cruz), anti-Nephrin (Santa Cruz), anti-cleaved caspase-3 (Cell signaling), anti-caspase-3 (Cell signaling) and anti- BAD (Cell signaling) antibodies at 4°C overnight. After washing, the membrane was incubated with horseradish peroxidase-conjugated secondary antibody (Jackson Labs). The reaction was visualized using an enhanced chemiluminescence system (Pierce). Immunoblots were analyzed by scanning densitometry and quantified by Quantity One gel Analysis software (Bio-Rad Laboratories).
Quantity one gel analysis software
Manufactured by Bio-Rad
Quantity One is a gel analysis software from Bio-Rad. It is designed to analyze and quantify bands or spots on electrophoresis gels, blots, and other types of gel-based images.
Automatically generated - may contain errors
Lab products found in correlation
Horseradish peroxidase conjugated secondary antibody, by Jackson ImmunoResearch (4 mentions)
Anti tsp 1, by Thermo Fisher Scientific (2 mentions)
Anti gapdh, by Merck Group (2 mentions)
Anti wt1, by Santa Cruz Biotechnology (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti nephrin, by Santa Cruz Biotechnology (1 mentions)
Anti bad, by Cell Signaling Technology (1 mentions)
Anti caspase 3, by Cell Signaling Technology (1 mentions)
Anti phospho gsk3β, by Cell Signaling Technology (1 mentions)
Anti gsk3β, by Cell Signaling Technology (1 mentions)
Anti sod, by Abcam (1 mentions)
Anti β actin, by Cell Signaling Technology (1 mentions)
Collagenase a, by Merck Group (1 mentions)
Anti phospho p38, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase, by Cell Signaling Technology (1 mentions)
Ab133303, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ripa lysis buffer, by Roche (1 mentions)
Anti cd47, by BD (1 mentions)
Anti nf κb p65, by Santa Cruz Biotechnology (1 mentions)
8 protocols using quantity one gel analysis software
1
Kidney Cortex Immunoblot Analysis
2
Assessing Bacterial Community Shifts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The compositional shifts of the bacterial communities were assessed by comparing the DGGE banding patterns of replicate salecan-treated samples to those of the control soils. The scanned banding patterns of the DGGE profiles were analyzed using Quantity One gel analysis software (Bio-Rad) to generate similarity dendrograms via the unweighted pair-group method using arithmetic averages (UPGMA) clustering method [33 ]. The results are expressed as the means ± standard deviation (SD). A P-value less than 0.05 was considered a statistically significant difference.
3
Immunoblot Analysis of INS-1 Cell Lysates
4
Isolation and Immunoblotting of Mouse Glomeruli
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glomeruli were isolated from mice by gradually sieving method as previously described [33 (link)]. Briefly, kidneys were minced and digested in collagenase A (Sigma) at 37°C for 30 minutes with gentle agitation. The digested kidney tissues were gently pressed through a 100-μm cell strainer. The filtered tissues were then passed through 70 μm and 40 μm cell strainer. The final filtered tissues were collected and centrifuged at 2,000 rpm for 5 minutes. The pellet enriched in glomeruli was collected. Mouse glomeruli and podocytes were homogenized. Equal amount of total protein was subjected to SDS-PAGE gel under reducing conditions and transferred onto a nitrocellulose membrane. After blocking, the membrane was incubated with anti-TSP1 (Thermo Scientific), anti-CD36 (Novua), anti-phospho-p38 and anti-total p38 antibodies (Cell Signaling) and then with horseradish peroxidase-conjugated secondary antibody (Jackson Labs). The reaction was visualized using an enhanced chemiluminescence system (Pierce). Immunoblots were analyzed by scanning densitometry and quantified by Quantity One gel Analysis software (Bio-Rad Laboratories).
5
Evaluating Smad3 Phosphorylation in Kidney
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kidney cortex was homogenized and equal amount of total protein was subjected to SDS-PAGE gel under reducing conditions and transferred onto nitrocellular membrane. After blocking, the membrane was incubated with anti-phospho-Smad3 or Smad3 antibodies (Cell Signaling) and then incubated with horseradish peroxidase-conjugated secondary antibody. The reaction was visualized using an enhanced chemiluminescence system (Pierce). Immunoblots were analyzed by scanning densitometry and quantified by Quantity One Gel Analysis software (Bio-Rad).
6
Western Blot Analysis of INS-1 Cell Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Following the corresponding treatments of the INS-1 cells, all cellular proteins were lysed in RIPA lysis buffer (Roche Diagnostics) containing protease inhibitors and the concentration was measured using a BCA protein assay kit (Beyotime Institute of Biotechnology, Shanghai, China). Total proteins (20–40 µg)were separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The separated proteins were then transferred onto a PVDF membrane followed by blocking the non-specific antigen and incubating with the corresponding primary antibody overnight. The primary antibodies used in this study were: a mouse anti-rat β-actin antibody (A5316; 1:20,000) and a rabbit anti-rat RAGE antibody (R5278; 1:1,000) (both from Sigma-Aldrich); a rabbit anti-rat PKCβ2 antibody (07-873-I; 1:1,000) and a mouse anti-rat TRB3 antibody (ST1032; 1:1,000) (both from Calbiochem, Billerica, MA, USA), and a rabbit anti-rat NOX4 antibody (ab133303; 1:1,000; Abcam). The secondary antibodies used in this study were a goat anti-mouse IgG antibody (A3682) and a goat anti-rabbit IgG antibody (A0545) (1:20,000; both from Sigma-Aldrich). An analysis of the protein bands was performed using Quantity One gel analysis software (Bio-Rad Laboratories, Inc.).
7
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Brow fat or skeletal muscle was homogenized in lysis buffer. After concentration being measured, homogenates were subjected to SDS-PAGE gel under reducing conditions and transferred onto a nitrocellulose membrane. After blocking, the membrane was incubated with anti-GAPDH (Millipore), anti-CD47 (BD Biosciences), or anti-PKG-I (BD Biosciences) antibodies at 4°C overnight. After washing, the membrane was incubated with horseradish peroxidase-conjugated secondary antibodies (Jackson Labs). The reaction was visualized by using an enhanced chemiluminescence system (Pierce). Immunoblots were analyzed by scanning densitometry and quantified by Quantity One gel Analysis software (Bio-Rad Laboratories).
8
Immunoblotting for Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein extraction and immunoblotting were performed as described in our earlier studies [43 (link),46 (link)]. In this study, the protein-transferred membranes were incubated with rabbit polyclonal anti-MYC, anti-Sox-2(Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA), anti-ABGC2, and anti-nCadherin (Aviva Systems Biology Corp., San Diego, CA, USA) antibodies, and mouse monoclonal anti-NFκB-p65, anti-NFκB-p50, anti-eCadherin (Santa Cruz), anti-NOTCH-1 (Pierce Biotechnology, Rockford, IL, USA), and mouse polyclonal CXCL12 (eBioscience Inc., San Diego, CA, USA). Membranes were developed with the appropriate anti-mouse/anti-rabbit (BioRad Laboratories, Hercules, CA, USA) secondary antibody. Blots were stripped and reblotted with rabbit polyclonal anti-β-actin antibody (Gentex Inc., Irvine, CA, USA) or anti-α-tubulin to determine equal loading of the samples. Densitometry analysis was performed using Quantity One gel analysis software (BioRad). α-tubulin or β-actin normalized values are compared between groups using t-test or two way ANOVA with Bonferoni’s post hoc correction (GraphPad Prism) and a P value of less than one is considered statistically significant.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!